LOCUS       Exported                6419 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMB18424, complete sequence.
ACCESSION   MG952432
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6419)
  AUTHORS   Batista MB.
  TITLE     pMB18424, a new tetA-sacB vector for counterselection in bacteria
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6419)
  AUTHORS   Batista MB.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-FEB-2018) Molecular Microbiology, John Innes Centre, 
            Colney Laney, Norwich, Norfolk NR4 7PH, United Kingdom
REFERENCE   3  (bases 1 to 6419)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6419
                     /organism="Cloning vector pMB18424"
                     /mol_type="other DNA"
                     /db_xref="taxon:2093648"
     CDS             complement(407..952)
                     /codon_start=1
                     /product="lacZ-alpha"
                     /label=lacZ-alpha
                     /protein_id="AVK78348.1"
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI
                     PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFAG
                     FPRQALNRGLPLGFRFSA"
     primer_bind     709..732
                     /label=M13 F(-47)
                     /note="M13 F(-47)"
     primer_bind     736..752
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        762..780
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    789..896
                     /label=MCS
                     /note="MCS"
     primer_bind     806..822
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(856..872)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(909..927)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(948..964)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(960..983)
                     /label=M13 R (-48)
                     /note="M13 R (-48)"
     regulatory      972..988
                     /regulatory_class="other"
                     /note="operator"
     protein_bind    972..988
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     regulatory      996..1026
                     /regulatory_class="promoter"
                     /note="lac promoter"
     promoter        complement(996..1026)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1041..1062
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(1221..2411)
                     /codon_start=1
                     /product="TetA"
                     /label=TetA
                     /protein_id="AVK78349.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     promoter        complement(2459..2487)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     promoter        2519..2964
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             2965..4386
                     /codon_start=1
                     /product="sacB"
                     /label=sacB
                     /protein_id="AVK78350.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVSEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     oriT            4705..4814
                     /note="incP origin of transfer"
     oriT            4752..4821
                     /label=RP4-oriT
                     /note="RP4-oriT"
     promoter        complement(5049..5139)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     promoter        complement(5333..5423)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     rep_origin      5783..6377
                     /label=pMB1 ori
                     /note="pMB1 ori"
     rep_origin      5783..6371
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"